The overall objective of the proposed research is to investigate the biosynthesis of a membrane protein and its structural and functional interaction with a membrane. The interaction of a defined, sequenced membrane associated protein, the fl bacteriophage coat protein, with the E. coli cytoplasmic membrane will be studied as a model system. The relative location of this protein in the cytoplasmic membrane will be investigated using various techniques such as group specific reagents, polyacrylamide gel electrophoresis, ferritin conjugated antibodies and electron microscopy. Interaction of the protein with various detergents and phospholipids will be studied to gain insight into the state of the coat protein in a lipid environment. The phospholipid biosynthetic enzymes will be investigated to determine the mechanism by which the presence of the coat protein in the cytoplasmic membrane alters the phospholipid synthesis and composition of the membrane. Attempts will be made to determine the intracellular site of biosynthesis of the coat protein and the role of phospholipid synthesis in the synthesis and insertion of the coat protein into the cytoplasmic membrane. Investigations will be carried out to establish what other effects insertion of the coat protein may have in altering other membrane and cell processes.